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P cofilin ser 3

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Sourced in Poland, United States

P-cofilin Ser 3 is a lab equipment product that detects the phosphorylation of cofilin at serine 3. Cofilin is an actin-binding protein that plays a role in the regulation of actin dynamics within cells.

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Lab products found in correlation

Cofilin, by Cell Signaling Technology (2 mentions) Gapdh, by Cell Signaling Technology (2 mentions) Cleaved caspase 3, by Cell Signaling Technology (1 mentions) Cleaved parp, by Cell Signaling Technology (1 mentions) Nf κb p65, by Cell Signaling Technology (1 mentions) E cadherin, by BD (1 mentions) β actin, by Santa Cruz Biotechnology (1 mentions) P src tyr416, by Cell Signaling Technology (1 mentions) Pfak tyr 925, by Cell Signaling Technology (1 mentions) Nf κb1 p105 p50, by Cell Signaling Technology (1 mentions) P iκbα ser32, by Cell Signaling Technology (1 mentions) P nf κb p65 ser536, by Cell Signaling Technology (1 mentions) Lamin a c, by Cell Signaling Technology (1 mentions) Snail, by Cell Signaling Technology (1 mentions) Las 4000 imager, by Fujifilm (1 mentions) Gapdh, by Thermo Fisher Scientific (1 mentions) Cleaved caspase 3, by Merck Group (1 mentions) Cleaved caspase 9, by Cell Signaling Technology (1 mentions) Supersignal west pico chemiluminescent substrate, by Thermo Fisher Scientific (1 mentions) P67phox, by Merck Group (1 mentions)

Spelling variants of this product

Cofilin (77 citations) P-cofilin (34 citations)

5 protocols using p cofilin ser 3

1

Investigating Focal Adhesion Kinase Signaling

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The p-FAK Tyr925 (cat #3284), FAK (cat #3285), p-src Tyr416 (cat #59548), src (cat #2109), p-cofilin Ser 3 (cat #3311), cofilin (cat #5175), NF-κB1 p105/p50 (cat #3035), NF-κB p65 (cat #8242), SNAIL (cat #3879), cleaved PARP (cat #5625), cleaved Caspase-3 (cat #9664), IκBα (cat #4814), Lamin A/C (cat #2032), p-NF-κB p65-Ser536 (cat #3033), and p-IκBα-Ser32 (cat #2859) antibodies were procured from Cell Signaling Technology and were used at 1:1000 dilution; β-actin (cat #sc-47778) antibody was from Santa Cruz Biotechnology and was used at 1:2000 dilution. E-cadherin (cat #610181) was from BD Biosciences and was used at 1:1000 dilution. MIEN1 antibody (cat #H00084299-M02) was from Abnova and was used at 1:2000 dilution. Human recombinant protein EGF was from Gibco (cat #PHG0311). MIEN1 protein was custom produced by Biomatik. The PCR primers were synthesized by Integrated DNA Technologies. The primer sequences are given in Table S2.
Tripathi A.K., Desai P.P., Tyagi A., Lampe J.B., Srivastava Y., Donkor M., Jones H.P., Dzyuba S.V., Crossley E., Williams N.S, & Vishwanatha J.K. (2024). Short peptides based on the conserved regions of MIEN1 protein exhibit anticancer activity by targeting the MIEN1 signaling pathway. The Journal of Biological Chemistry, 300(3), 105680.
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2

Caspase Activation Analysis by Western Blot

Cited 1 time
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Following treatment with desired drugs, attached cells were harvested and analyzed for caspase activation by Western blot analysis as previously described [11] (link). The blots were then probed with primary antibodies to ROCK1 (GTX61382) from GeneTex, ROCK2 (sc-5561) from Santa Cruz Biotechnology, p67phox (07-502) from Millipore, cleaved caspase 3 (#9661), cleaved caspase 9 (#9509), cleaved caspase 8 (#9429), PARP (#9542), cofilin (#3312), p-cofilin(Ser3) (#3311), MLC (#3672), p-MLC(Ser19) (#3671) from Cell Signaling. After blotting with corresponding secondary antibodies conjugated with horseradish peroxidase, the membranes were developed with ECL Western blotting or SuperSignal West Pico Chemiluminescent Substrate (Thermo Scientific), and the blots were visualized by using a Fujifilm LAS-4000 Imager. All blots were normalized to GAPDH (MA5-15738, Thermo Scientific) or to actin (sc-1616; Santa Cruz Biotechnology).
Surma M., Handy C., Chang J., Kapur R., Wei L, & Shi J. (2014). ROCK1 Deficiency Enhances Protective Effects of Antioxidants against Apoptosis and Cell Detachment. PLoS ONE, 9(3), e90758.
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3

Protein Expression Analysis via Western Blot

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The cells were washed with cold PBS buffer and total proteins were extracted from the collected cells using RIPA buffer (25 mM Tris–HCl (pH 7.4), 150 mM NaCl, 0.1% SDS, 0.5% sodium deoxycholate, 1mM EDTA, 1% Nonidet P-40) and protease inhibitors. The bicinchoninic acid (BCA) assay was performed to determine the total protein concentration in the samples (Pierce, Rockford, IL, USA). Samples were fractionated on a 4–20% SDS-PAGE gels. Next, the proteins were transferred onto a nitrocellulose membrane and probed with anti-human/mouse antibodies specific to PARP (#9542), caspase 3 (#9668), p-BAD Ser112 (#9291), p-cRaf Ser338 (#9427), p-ERK1/2 Thr202/Tyr204(#4370), p-PAK1 Thr423/PAK2 Thr402 (#2601), PAK2 (2608), p-cofilin Ser3 (#3313), γH2AX (#9718), p-STAT3 Tyr705 (#9138), and GAPDH (#5174) (purchased from Cell Signaling Technologies, LabJOT, Warsaw, Poland). Signals were detected using the enhanced chemiluminescence detection system (MicroChemi Bio-Imaging Systems). The images of row Western blot membranes are shown in Figure S2.
Flis S., Bratek E., Chojnacki T., Piskorek M, & Skorski T. (2019). Simultaneous Inhibition of BCR-ABL1 Tyrosine Kinase and PAK1/2 Serine/Threonine Kinase Exerts Synergistic Effect against Chronic Myeloid Leukemia Cells. Cancers, 11(10), 1544.
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4

Quantifying Cytoskeletal Protein Dynamics

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Total protein lysates were generated using RIPA buffer (Sigma Aldrich, St. Louis, MO, USA) according to manufacturer’s instructions, supplemented with protease inhibitors, and separated on 10% TGX gel (Bio-Rad, Hercules, CA, USA) under reducing conditions. The gel was transferred to a PVDF membrane and probed with 1:3000 class III β-tubulin (TUJ1; BioLegend, San Diego, CA, USA), 1:10,000 GAPDH (Cell Signaling Technologies, Danvers, MA, USA), Cofilin (D3F9) (1:1000, Cell Signaling Technologies), p-Cofilin (Ser3) (1:1000, Cell Signaling Technologies), monoclonal α-tubulin (DM1A) (1:2000, Sigma Aldrich), anti-detyrosinated α-tubulin (glu-tubulin) (1:1000, Abcam, Waltham, MA, USA), tyrosinated α-tubulin (1:2000, Sigma Aldrich), acetyl α-tubulin (Lys40, D20G3) (1:1000, Cell Signaling Technologies), α-Actin (AC-15) (1:10,000, Sigma Aldrich, Burlington, MA, USA), CAPZB (1:1000, Bio-Rad), and Diap1 (1:1000, Cell Signaling Technologies). Standard molecular weight markers were used (Precious Plus Protein Standards #1610374/ #1610375, BioRad, St. Louis, MO, USA). Densitometry was analyzed with ImageJ: https://imagej.nih.gov/ij/ (accessed on 11 June 2018). Experiments were performed in duplicate and representative images are shown. Raw western blot figures can be found in the supplementary materials.
Reader J.C., Fan C., Ory E.C., Ju J., Lee R., Vitolo M.I., Smith P., Wu S., Ching M.M., Asiedu E.B., Jewell C.M., Rao G.G., Fulton A., Webb T.J., Yang P., Santin A.D., Huang H.C., Martin S.S, & Roque D.M. (2022). Microtentacle Formation in Ovarian Carcinoma. Cancers, 14(3), 800.
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5

Protein Extraction and Analysis from Mouse Aortas

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Mouse aortas were harvested, cleaned of fat and connective tissue, and then flash frozen in liquid nitrogen. Frozen aortas were ground into a fine powder using a pre-chilled mortar and pestle. Proteins were extracted in Hunters buffer (25 mmol/L HEPES, 150 mmol/L KCl, 1.5 mmol/L MgCl2, 1 mmol/L EGTA, 10 mmol/L Na-pyrophosphate, 10 mmol/L NaF, 1% Na deoxycholate, 1% Triton X 100, 0.1% SDS, 10% Glycerol, Na-orthovanadate and protease inhibitors) from there lysates sonicated and cleared at 10,000 x g for 5 minutes. Proteins were separated using SDS-PAGE and transferred to Immobilon-P polyvinylidene difluoride (PVDF) membranes (Millipore), blocked with 5% non-fat dairy milk, and incubated with primary antibodies to SSH1 (Cell Signaling Cat #13578), p-cofilin ser-3 (Cell signaling Cat #3311), cofilin (Gentex Cat# GTX102156), OPN (R&D # AF808), TGFβ1 (Cell Signaling #3711), TGFβRII (Santa Cruz Cat# sc-17799) smooth muscle 22 (SM22) (Abcam 14106) or β-actin (Sigma Cat #A5441). Subsequently, blots were incubated with horseradish peroxidase-conjugated secondary antibodies and proteins were detected by enhanced chemiluminescence (ECL, Millipore). Images were acquired using a Carestream Molecular Imaging (Carestream) device. Band intensity was quantified by densitometry using Image J software (NIH).
Williams H.C., Ma J., Weiss D., Lassègue B., Sutliff R, & Martín A.S. (2018). The cofilin phosphatase slingshot homolog 1 restrains angiotensin II-induced vascular hypertrophy and fibrosis in vivo. Laboratory investigation; a journal of technical methods and pathology, 99(3), 399-410.
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